(精选)【医学课件课件】结直肠癌变各阶段的全基因组外显子测序结合数字化表达谱研究教学课件.pptVIP

结直肠癌变各阶段的全基因组外显子测序结合数字化表达谱研究 杨柳 浙江大学肿瘤研究所 浙江大学教育部肿瘤预警与干预重点实验室 肿瘤的形成过程发生了一系列基因组遗传变异，包括体细胞突变、基因组结构变异和拷贝数改变等。 研究背景 NATURE|Vol 458|9 April 2009 结直肠独特的病理过程（正常粘膜－腺瘤－腺癌）为肿瘤癌变过程研究提供了一个非常好的模型。 新一代测序技术的发展，从全基因组水平来分析结肠肿瘤癌变各阶段的基因谱图成为可能。 PNAS March 18, 2008 vol. 105 no. 11 4283–4288 研究方法： 正常肠粘膜组织 腺瘤 腺癌 提取DNA、RNA NimbleGen 2.1M Human Exome Array芯片捕获全基因组外显子 Illumina GA II 测序仪高通量测序 数字表达谱分析 数据分析：单核苷酸变异，小片断插入、缺失 ，突变基因GO/PATHWAY，差异基因 Exome Capture Sequencing N: 正常上皮 A: 腺瘤 T: 腺癌 Male, 66y Grading IIB, Adenocarcinoma Negative family cancer history 研究材料： Summary of Exome-capture Data 　 Normal mucosa Adenoma Carcinoma Average sequencing depth on target 23.00 60.48 58.20 Raw Data (Gb) 2.76 6.94 6.93 Effective sequence on or near target (Gb) 1.65 4.27 4.19 Total effective reads (Gb) 29.65 74.03 75.40 Effective sequence on target (Gb) 0.77 2.01 1.94 Coverage of target region 98.00% 99.00% 98.90% Summary of SNV (directions) identified in adenoma and carcinoma of one CRC patient 　 Adenoma versus normal mucosa Carcinoma versus normal mucosa Total Total SNVs 445 720 1073 Synonymous 117 301 384 Missense 320 410 673 Nonsense 3 5 8 Readthrough 5 4 8 SNVs-Genes 277 321 533 Summary of Indels identified in adenoma and carcinoma of one CRC patient 　 Adenoma versus normal mucosa Carcinoma versus normal mucosa Total Indels 1043 843 1452 CDS 352 298 483 Intron 576 451 796 3’-UTR 20 20 34 5’-UTR 16 9 21 Others 79 70 118 Mutation tendency of SNV in the sequential process of normal mucosa-adenoma-carcinoma (N-A-C) Non-synonymous mutation-Genes in adenoma and carcinoma. Go analysis of common mutations genes in adenoma and carcinoma. Pathway analysis of common mutations genes in adenoma and carcinoma. KEGG pathway distribution * *